Quantitative ELISA for mouse monoclonal antibody determination in culture supernatants and in human serum.
An ELISA has been set up for quantifying mouse monoclonal antibodies in culture supernatant. The assay includes rabbit anti-mouse IgG antibodies chromatographycally purified. This preparation was used as coating and as conjugated antibodies in the ELISA. The assay can detect IgG1 with sensitivity of 0.2 ng/mL, IgG2a (0.85 ng/mL), IgG2b (0.13 ng/mL), and IgG3 (3.19 ng/mL) in culture supernatants. The effective working range was from subnanogram per mL quantities to 30 ng/mL by using a computer statistical program. Variation coefficient of ELISA was below 7%. Correlation estimates with a similar ELISA using commercial reagents were performed for each mouse antibody subclass. The assay was able to detect the four mouse monoclonal antibody subclasses in pure human serum as compared with the same ELISA using commercial antibodies. A 24-h pharmacokinetic profile of 1 patient treated with an IgG2a monoclonal antibody is presented.
Vilaseca
J C
JC
Institute of Tropical Medicine Pedro Kouri, Ciudad de la Habana, Cuba.
Pupo
M
M
Bernal
M
M
Matamoros
L
L
Gordillo
S
S
Rodríguez
H
H
Caballero
Y
Y
Otero
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D051379:Mice
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G